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1. Would the materials used in your project and/or your final product pose:

Risks to the safety and health of team members or others in the lab?

Working in the lab always has risks for people but we will take action to minimize any risk, some of the precautions we take are listed here: • The microorganisms will be grown in petri dishes, sealed completely during bacterial growth, thus preventing their escape into stages of experimentation. • Inoculation experiments will be conducted on plants that are isolated in a greenhouse. Plants will be kept for a minimum time of experimentation and will be discarded as biohazard material immediately after the experiments. • Equipment such as biological safety cabinet class I and laminar flow cabinet will be used for minimize the possibility of spread of microorganisms. • All students involved with the project will be informed of the established security arrangement and sign a commitment to strictly comply the rules. • Also, prior to the beginning of the experimental procedures, all of the team members were given a course of biosafety, focused mainly on biological hazards, chemical reagents and general BSL-1 laboratory safety concerns and security procedures.

The system design tests will be conducted within a laboratory of biosafety level 1, and we will use the following materials and microorganisms strains:

Bacterial strains:

Escherichia coli K12

Is classified as a non-pathogenic microorganism for plant, animal or man. The genetic manipulations that we will be making are routinely in most academic laboratories in the world. For more information visit [http://epa.gov/biotech_rule/pubs/fra/fra004.htm Escherichia coli]

Known toxic chemical reagents and hazardous physical agents:

Ethidium bromide

An intercalating agent, toxic and suspected mutagenic, is used widely used as a nucleic acid fluorescent stain in molecular biology, in our project Ethidium bromide will be used only in electrophoretic procedures. Some of the precautions we take when working with this mutagenic agent are always wearing nitrile gloves and working in a delimitated area, avoiding direct skin contact an inhalation, specially isolated to prevent involuntary exposure of laboratory coworkers with Ethidium bromide.

Ultraviolet light

Is an electromagnetic radiation with a wavelenght shorter that of visible light. We will use UV light in the biological safety and laminar flow cabinets for sterilization and decontamination purposes; also, UV-light would be used for visualization of the stained DNA in the electrophoretic gels.

In order to prevent accidental exposition to UV radiation, precautions would be taken, and when necessary special safety lenses would be used, also we will always use UV-blocking shields when visualizing electrophoretic gels in UV light.

All other toxic and chemicals will be handled to avoid direct contact, and observing the proper safety procedures, additionally, all chemicals reagents and biological materials will be disposed only in the designated biohazard receptacles, following the regulations of the University for [http://departamentomedico.uniandes.edu.co//Documentos/Salud_Ocupacional/Procedimientomanejoresiduossolidos.pdf Guidelines for Integral Solid Waste Management] of the Medical and Occupational Health Department at the Universidad de los Andes.

Risks to the safety and health of the general public if released by design or accident?

It is important to mention that, although our project proposes an agricultural application of the strains generated, in this stage of the investigation we will not do toxicity tests in animals or humans of the genetically modified organism. The risks in an accident are minimal because the Escherichia coli K12 strain is not pathogenic for humans, and the genetic modifications that we will make do not improve the ability of the bacteria to prejudice the human health.

Is also important to mention that no human nor animal pathogenic strains of bacteria are going to be used in the current research; nevertheless the national directions regarding Genetically Engineered Organisms (GMOs) will be followed.

Risks to environmental quality if released by design or accident?

Although the initial stages of our research are mainly laboratory-based, the goal is to implement our system directly in coffee plantations. Taken this into account, we’re planning to develop an strategy to build our bacterial-based detection system on organisms representing low-to-no risk for the normal microbiota of the plantations and innocuous for human consumption.

We will not release the genetically modified organism to the countryside. In case of accident the Escherichia coli k12 strain is no pathogenic for animals or plants, and the genetic modifications are not an advantage to damage the environmental quality

Risks to security through malicious use by individuals, groups or states?

Taking into account that the genetic modifications we will make to the Escherichia coli k12 strain are focused to improve the ability of plants to survive a fungi infection and that these modifications do not have any danger to people, plants, animals, or environment, then our project does not have any risk to security through malicious use by individuals, groups or states.